LOCUS       sequence-358262-        8062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MYC-tagged Acanthoeca spectabilis NF-kB1 protein in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8062)
  AUTHORS   Williams LM, Sridhar S, Samaroo J, Peart J, Adindu EK, Addanki A, 
            DiRusso CJ, Aguirre Carrion PJ, Rodriguez-Sastre N, Siggers T, 
            Gilmore TD
  TITLE     Comparison of NF-kappaB from the protists Capsaspora owczarzaki and 
            Acanthoeca spectabilis reveals extensive evolutionary 
            diversification of this transcription factor.
  JOURNAL   Commun Biol. 2021 Dec 16;4(1):1404. doi: 10.1038/s42003-021-02924-2.
  PUBMED    34916615
REFERENCE   2  (bases 1 to 8062)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2021-12-16"; volume: "4(1):1404. doi"; pages: " 
            10.1038/s42003-021-02924-2"
COMMENT     Sequence Label: pcDNA-MYC-As-NF-kB1
FEATURES             Location/Qualifiers
     source          1..8062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3..32
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             42..71
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             81..110
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             120..149
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             159..188
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             222..251
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2719..2736)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2725..2949
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2995..3423
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3082..3101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3292..3313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3432..3452)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3437..3766
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3617..3752
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3679..3698
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3833..4627
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3887..3906)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4497..4516
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4801..4922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4838..4857)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4892..4911
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4971..4987)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4971..4987)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4984..5006)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4995..5011
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5019..5025,5026..5043,5044..5049))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5064..5085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5202..5219)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5373..5961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5453..5472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6132..6923,6924..6992))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6755..6774
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6993..7097)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7172..7191)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7363..7742
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7743..7946
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7897..7917
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7991..8010
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7991..8009
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tggagcaaaa gctcatttct gaagaggact tgaatgaaat ggagcaaaag ctcatttctg
       61 aagaggactt gaatgaaatg gagcaaaagc tcatttctga agaggacttg aatgaaatgg
      121 agcaaaagct catttctgaa gaggacttga atgaaatgga gcaaaagctc atttctgaag
      181 aggacttgaa tgaaatggag agcttgggcg acctcaccat ggagcaaaag ctcatttctg
      241 aagaggactt gaattcttac aagatggctt ctggagagca ccgaaattta gttcgcagag
      301 gtgcacccat gattcctgaa cacagagcgg ctttgctcga aaaatgccac aaatctgaac
      361 ctccacaagg gaaagatttt tacgattcag gctttggggg tagtttagaa gaagaggcag
      421 atatttttga acgtgaacaa gataagcttg tggaagcaac gagggaaata acgcttgatg
      481 aatcaccgat atatggactc gacaagaaag aaactgaaca acctcagtgt acgcctccgc
      541 ctcaaatgga aagtgatcct atttcagacc ccgaaatttc gcagagtcag tcgatttcac
      601 cccataattg tgtgctaagc tacgcagctg gtaatgatgt gcgatgcttg cttgctccta
      661 tgaggggctt gttgttcgct caagacgaag atggcgatac tgcccttcat ctggccataa
      721 tccacactaa cgtccaagcg gtggaaaaca ttgtcgcagc tgcaccatct acgaaagctc
      781 ttgatatttt caactacttg agacaaaccc cactacacct tgcaaccatc accaaacagt
      841 caaatattgt tcgtgggctg attgcatctg gtgcaagtgt agacctggtg gatagaaatg
      901 gaaaaaccgc cctccatctc gcatgcgaaa gaggtgacat agacagtgtg cgcgagatta
      961 taaggccatt atcagataag gcatacaacc ccaagaccag ggaagagata tcctccatct
     1021 tgaatactcg caactatgac ggcttcacag cacttcatgt tgccgtattt tccaactcta
     1081 ttgacattgt gagcgcactc acaaatgttg gagcagacat caatgtccct gattgcggaa
     1141 gtggtaagac agcattacac catgccgtag aaaccaacaa cctacgctta gtcagctact
     1201 tactgttcca gtgcaatgct tgcgtagatg ccgaaacatt tgatgaatgc acaccgctgc
     1261 attttgctgc tgggcgtggc atggagtcta tggctgcatt actactggcg gcaggcgccg
     1321 acccaacact gcccaacagg actggagcca caccgcttga tgaagccgct gataatgtac
     1381 ggccaatgct ccagctaccc gaatcaccct tctagacaga gaattccatg aataggcacc
     1441 tctggaagag ccaactgtgt gagatggtgc agcccagtgg tggcccggca gcagatcagg
     1501 acgtactggg cgaagagtct cctctgggga agccagccat gctgcacctg ccttcagaac
     1561 agggcgctcc tgagaccctc cagcgctgcc tggaggagaa tcaagagctc cgagatgcca
     1621 tccggcagag caaccagatt ctgcgggagc gctgcgagga gcttctgcat ttccaagcca
     1681 gccagaggga ggagaaggag ttcctcatgt gcaagttcca ggaggccagg aaactggtgg
     1741 agagactcgg cctggagaag ctcgatctga agaggcagaa ggagcaggct ctgcgggagg
     1801 tggagcacct gaagagatgc cagcagcaga tggctgagga caaggcctct gtgaaagccc
     1861 aggtgacgtc cttgctcggg gagctgcagg agagccagag tcgcttggag gctgccacta
     1921 aggaatgcca ggctctggag ggtcgggccc gggcggccag cgagcaggcg cggcagctgg
     1981 agagtgagcg cgaggcgctg cagcagcagc acagcgtgca ggtggaccag ctgcgcatgc
     2041 agggccagag cgtggaggcc gcgctccgca tggagcgcca ggccgcctcg gaggagaaga
     2101 ggaagctggc ccagttgcag gtggcctatc accagctctt ccaagaatac gacaaccaca
     2161 tcaagagcag cgtggtgggc agtgagcgga agcgaggaat gcagctggaa gatctcaaac
     2221 agcagctcca gcaggccgag gaggccctgg tggccaaaca ggaggtgatc gataagctga
     2281 aggaggaggc cgagcagcac aagattgtga tggagaccgt tccggtgctg aaggcccagg
     2341 cggatatcta caaggcggac ttccaggctg agaggcaggc ccgggagaag ctggccgaga
     2401 agaaggagct cctgcaggag cagctggagc agctgcagag ggagtacagc aaactgaagg
     2461 ccagctgtca ggagtcggcc aggatcgagg acatgaggaa gcggcatgtc gaggtctccc
     2521 aggccccctt gccccccgcc cctgcctacc tctcctctcc cctggccctg cccagccaga
     2581 ggaggagccc ccccgaggag ccacctgact tctgctgtcc caagtgccag tatcaggccc
     2641 ctgatatgga caccctgcag atacatgtca tggagtgcat tgagtagtct agagggcccg
     2701 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2761 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2821 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2881 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2941 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3001 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3061 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3121 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3181 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3241 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3301 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3361 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3421 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3481 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3541 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3601 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3661 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3721 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3781 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3841 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3901 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3961 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4021 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4081 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4141 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4201 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4261 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4321 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4381 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4441 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4501 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4561 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4621 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4681 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4741 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4801 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4861 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4921 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4981 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5041 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5101 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5161 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5221 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5281 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5341 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5401 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5461 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5521 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5581 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5641 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5701 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5761 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5821 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5881 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5941 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6001 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6061 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6121 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6181 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6241 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6301 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6361 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6421 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6481 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6541 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6601 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6661 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6721 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6781 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6841 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6901 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6961 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7021 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7081 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     7141 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7201 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     7261 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     7321 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     7381 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     7441 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     7501 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     7561 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     7621 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     7681 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     7741 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     7801 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     7861 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     7921 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
     7981 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt ttaaacttaa
     8041 gcttggtacc gatttaaagc ta
//
